In the present STTR we propose to investigate two novel approaches in our exploration of the therapeutic and regenerative potential of cell-based medicine. Furthermore, we will utilize a preclinical animal model of compression syndrome (CS) that mimics the pathophysiology of multiple injuries such as crush injuries, burns, circulation blockage, fractures, hemorrhage, severely bruised muscles, surgery and other trauma which lead to devastating tissue damage with potential loss of limb or life. These injuries result in damage to the muscular, neural and vascular components of the skeletal muscle compartment. We will examine amniotic fluid-derived (AFS) stem cells in combination with ASC101 in this small animal CS model. The important features of AFS cells that underscore their therapeutic potential include (1) ability to be expanded to a greater extent than other adult stem cells, (2) anti-inflammatory and (3) have multipotential differentiation capabilities but (4) do not show signs of malignant transformation in culture or form teratomas. The development of these novel stem cells has been pioneered by Dr. Soker and his colleagues at the Wake Forest Institute of Regenerative Medicine for the past 10 years. ASC101 is a new technology developed by America Stem Cell (ASC). ASC has shown the ability for ASC101 treatment to enhance selectin-mediated homing of specific stem cells to the bone marrow and ischemic tissue bed. Use of this technology involves a simple incubation of the stem cells with ASC101 just prior to injection. Our presently proposed studies are divided into two Specific Aims (SA). In SA#1 we will identify the parameters required for optimal delivery of AFS cells to the damaged tissue and assess whether ASC101 enhances cell homing. These parameters include time of injection post injury, optimal dose and residency time of the injected stem cells in the damaged tissue. In SA#2 we will utilize these parameters in a CS model that mimics the pathophysiology of multiple injuries. We will conduct not only anatomical analyses of various cell populations in the injured area post trauma but also functional tests to assess muscle functional recovery. The successful implementation of ASC101 with AFS cells to treat injuries associated with CS would be directly relevant to improving our understanding of the muscle recovery processes that occur following multiple types of injuries. In addition, the combination of ASC101 with AFS cells could enhance the therapeutic and regenerative capacity of these cells and most importantly provide an off-the shelf, effective solution for tissue damage due to multiple types of injuries or diseases. PUBLIC HEALTH RELEVANCE: A variety of injuries, such as crush injuries, burns, circulation blockage, fractures, hemorrhage, severely bruised muscles, surgery and other trauma lead to devastating tissue damage with potential loss of limb or life. To address the complexity of the events underlying the pathophysiology of these injuries we will explore the therapeutic potential of a cell-based approach. We will utilize a novel type of stem cell exhibiting multilineage differentiation potential in combination with a novel technology that can enhance the migration of cells to the site of damage. The potential for broad therapeutic application of this combination approach underscores the importance of these proposed studies.